DESCRIPTION (APPLICANT'S ABSTRACT): N-Acylethanolamines (NAEs) are potent, bioactive lipid mediators with diverse physiological roles in mammals including, but not restricted to, the control of pain initiation. Anandamide, a type of NAE with arachidonic acid as the fatty acid constituent, is an endogenous ligand for the cannabinoid receptor (CBI type) in mammalian brain, and the transient extracellular accumulation of anandamide (and other NAEs) is part of the "endocannabinoid" signaling pathway in neuronal cells involved in the perception of pain. NAEs recently were found to be enriched in seeds of higher plants, and we propose in this exploratory research and development phase to identify abundant, natural sources of NAE-mixtures (combined with other endocannabinoid lipid mediators such as acylglycerols) which could be used in the development of "natural analgesics." In support of this application, accurate quantitative procedures (gas chromatography coupled with mass spectrometry) were developed to identify these lipid metabolites in crude plant extracts and in industrially-refined vegetable oil fractions. Moreover, a rapid screening method is in place by which various extracts can be tested for their neurological activities in vitro. This method relies on quantitative multi-channel electrophysiological measurements of action potential patterns recorded from neural networks cultured on microelectrode arrays (MEAs). The reproducible "quieting" effects of several endocannabinoids (including a seed extract of mixed NAE species) on spontaneous and bicuculline-stimulated neural network activity have helped to verify the efficacy of this system. CBI-receptor-dependent and -independent mechanisms will be identified by classical pharmacological approaches (agonists, antagonists, inhibitors, etc.) as well as by molecular genetic approaches (cultures derived from CBI-knockout mice). Results from this research should target new botanical sources for development as neuroactive therapeutics.